Dear Flowers and flowerettes, how do you check your new batches of PMA (to activate cells to produce cytokines for intracellular staining)?? I thought I could check if the CD4 disappeared so I incubated my whole blood for 4 hours with the usual mixture (PMA/ionomycin/monensin). After surface staining the CD4 was still on the cells, but much dimmer respect to the control (monensin only). Lets say one log less. This is the first batch of PMA I buy here and I just started to test the anti-cytokines antibodies, so I don't have previous results to compare to. I also work on sheep blood, so I don't have access to fancy human/mouse reagents and controls. Do you expect a complete disappearance of your CD4 signal? And do you think 4 hours is too early to see a complete downregulation? I am considering to re-titrate my PMA and to judge the results on the basis of the CD4 downregulation. Do you think that is the right approach or am I assuming something completely wrong (CD4 downregulation directly associated to activation)? Any help will be appreciated. Thanks, Mara *********************** Mara S.L. Rocchi Moredun Research Institute International Research Centre Pentland Science Park Bush Loan Penicuik EH26 0PZ Scotland UK e-mail: roccm@mf.mri.sari.ac.uk Tel: 0131-4455111 ext. 464/461 Fax: 0131-446235 "The information contained in this email is confidential. It may also be legally privileged. It is intended only for the stated addressee(s) and access to it by any other person is unauthorised. If you are not an addressee, you must not disclose, copy, circulate or in any other way use or rely on the information contained in this email. Such unauthorised use may be unlawful. If you have received this email in error please reply to it immediately and delete it and all copies from your system."
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:26 EST